Immunodiagnosis and Immunotherapy of Tetanus and Botulinum Neurotoxins

  • J. L. Middlebrook
  • J. E. Brown
Part of the Current Topics in Microbiology and Immunology book series (CT MICROBIOLOGY, volume 195)


Tetanus and botulism are diseases or poisonings that are highly dramatic in their presentations. They are almost always life threatening and therefore command attention and concern by those around the victim. In earlier times help might have been sought from the shaman or witch doctor, whose spells or potions provided little comfort from the symptoms and eventual death. As modern medicine developed in the last two centuries even well-trained physicians were unable to do much more than a shaman until the advent of immunotherapy in the late 1800s and immunization in the 1920s. Little has changed since then, and we still depend on active immunity or passive immunotherapy to treat both of these highly dangerous poisonings. Immunological techniques and reagents also provide the means of toxin identification, and a timely and definitive diagnosis therefore depends heavily on their use. This chapter reviews the past and current literature on vaccines and various antibody-based products for treatment or prevention of poisonings by both botulinum and tetanus neurotoxins. We also review the techniques employed for their detection and identification.


Botulinum Toxin Immune Globulin Tetanus Toxoid Tetanus Toxin Clostridium Botulinum 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


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  1. Arnon SS (1993) Clinical trial of human botulism immune globulin. In: DasGupta BR (ed) Botulinum and tetanus neurotoxins, neurotransmission and biomedical aspects. Plenum, New York, pp 477–482Google Scholar
  2. Arunachalam B, Ghosh S, Talwar GP, Raghupathy R (1992) A single monoclonal antibody that confers total protection from tetanus. Hybridoma 11: 165–179PubMedGoogle Scholar
  3. Bardenwarper HW (1962) Serum neuritis from tetanus antitoxin. JAMA 179: 763–766Google Scholar
  4. Barile M, Hardegree M, Pitmann (1970) Immunization against neonatal tetanus in New Guinea III. The toxin-neutralization test and the response of guinea pigs to the toxoids as used in the immunization schedules in New Guinea. Bull World Health Organ 43: 453–459PubMedGoogle Scholar
  5. Barington T, Gyhrs A, Kristensen K, Heilmann C (1994) Opposite effects of actively and passively acquired immunity to the carrier on responses of human infants to a Haemophilus influenzae type b conjugate vaccine. Infect Immun 62: 9–14PubMedGoogle Scholar
  6. Bengtson IA (1921) Standardization of botulism antitoxins. Am J Public Health 11: 352–357Google Scholar
  7. Bengtson IA (1924) Studies on organisms concerned as causative factors in botulism. Bull US Hyg Lab 136: 1–101Google Scholar
  8. Beytout J, Lauras H, Cailleba A, Nguyen TT (1989) Rapid evaluation of tetanus immunity by a haemagglutination test in the injured at a hospital emergency unit. Biomed Pharmacother 43: 621–625PubMedGoogle Scholar
  9. Biglan AW, Gonnering R, Lockhart LB, Rabin B, Fuerste FH (1986) Absence of antibody production in patients treated with botulinum A toxin. Am J Ophthalmol 101: 232–235PubMedGoogle Scholar
  10. Bizzini B (1993) Clostridium tetani. In: Gyles C, Thoen C (eds) Pathogenesis of bacterial infections in animals, 2nd edn. Iowa State University Press, Ames, pp 97–105Google Scholar
  11. Bizzini B, Leonardi MS, Zummo S, Mastroeni P (1990) Screening of tetanus antibodies with the aid of dot-enzyme-linked immunosorbent assay (dot-ELISA). Validation of the results using standard ELISA and passive hemagglutination test. Boll Ist Sieroter Milan. 69: 391–393PubMedGoogle Scholar
  12. Black RE, Gunn RA (1980) Hypersensitivity reactions associated with botulinal antitoxin. AM J Med 69: 567–570PubMedGoogle Scholar
  13. Bleck TP (1991) Tetanus: pathophysiology, management, and prophylaxis. Dis Mon 37: 545–603PubMedGoogle Scholar
  14. Bolotovskii VM, Kriukov V, Raupov BE (1992) A comparative evaluation of the effectiveness of methods for serological sampling examinations. Zh Mikrobiol Epidemiol Immunobiol 1992: 23–26Google Scholar
  15. Bowmer EJ (1962) Antitoxins of Clostridium botulinum type A,B,C,D and E. Preparation and assay of proposed international standards. Thesis, University of LiverpoolGoogle Scholar
  16. Bowmer EJ (1963) Preparation and assay of the international standards for Clostridium botulinum types A,B,C,D and E antitoxins. Bull World Health Organ 29: 701–709Google Scholar
  17. Boyd J (1959) Symposium of tetanus. Tetanus in two world wars. Proc R Soc Med 52: 109PubMedGoogle Scholar
  18. Brin MF, Fahn S, Moskowitz C, Friedman A, Shale HM, Greene PE, Blitzer A, List T, Lange D, Lovelace RE, McMahon D (1987) Localized injections of botulinum toxin for the treatment of focal dystonia and hemifacial spasm. Mov Disord 2: 237–254PubMedGoogle Scholar
  19. Brown JE, Taylor KL, Brown WL, Clayton MA, Schmidt JJ, Jaax GP, Franz DR (1991) Hyperimmunization of horses for production of immune globulin to botulinum toxins. Abstracts, 10th world congress on animal, plant, and microbial toxins, Venom and Toxin Research Group, University of Singapore, Singapore, p 95Google Scholar
  20. Brussow H, Sidoti J, Freire WB (1993) Tetanus and diphtheria immunization coverage in Ecuadorian children after a national vaccination campaign. J Infect Dis 168: 479–483PubMedGoogle Scholar
  21. Cardella MA (1964) Botulinum toxoids. In: Lewis KH, Cassel KJ (eds) Botulism. U.S. department of health, education, and welfare, Cincinnati, pp 113–130Google Scholar
  22. Charles IG, Rodgers BC, Makoff AJ, Chatfield SN, Slater DE, Fairweather NF (1991) Synthesis of tetanus toxin fragment C in insect cells by use of a baculovirus expression system. Infect Immun 59: 1627–1632PubMedGoogle Scholar
  23. Chatfield SN, Fairweather N, Charles I, Pickard D, Levine M, Hone D, Posada M, Strugnell RA, Dougan G (1992) Construction of a genetically defined Salmonella typhi Ty2 aroA, aroC mutant for the engineering of a candidate oral typhoid-tetanus vaccine. Vaccine 10: 53–60PubMedGoogle Scholar
  24. Cilia EG, Saenz DJ, Urbieta EM, Carcedo MA, Perez TE (1993) Immunity against tetanus in childhood. An Esp Pediatr 38: 499–502Google Scholar
  25. Clare JJ, Rayment FB, Ballantine SP, Sreekrishna K, Romanos MA (1991) High-level expression of tetanus toxin fragment C in Pichia pastoris strains containing multiple tandem integrations of the gene. Biotechnol 9: 455–460Google Scholar
  26. Cooper MS (1964) Antitoxins to C. botulinum. In: Lewis KH, Cassel KJ (eds) Botulism. U.S. department of health, education, and welfare, Cincinnati, 147–164Google Scholar
  27. Council of Europe (1981, 1985) European Pharmacopoeia, 2nd edn. Maisonneuve, Saint-RuffinelGoogle Scholar
  28. Craven CJ, Dawson DJ (1973) The chain composition of tetanus toxin. Biochim Biophys Acta 317: 277–285PubMedGoogle Scholar
  29. D’Amelio R, Stroffolini T, Wirz M, Biselli R, Matricardi PM, Gentili G, Collotti C, Pasquini P (1993) Tetanus immunity among Italians born after the introduction of mandatory vaccination of children. Vaccine 11: 935–936PubMedGoogle Scholar
  30. D’Argenio P, Wirz M, Rosmini F, Adamo B, Gentili G, Puccinelli M, Collotti C, Mele A, Esposito D, Forte P, Pasquini P (1992) Tetanus immunity in a partially vaccinated population of children in a district of Naples (Secondiglianio). Eur J Epidemiol 8: 93–97PubMedGoogle Scholar
  31. Dastur FD, Shastry P, Iyer E, Awatramani V, Raut S, Mehta SD, Irani SF (1993) The foetal immune response to maternal tetanus toxoid immunization. J Assoc Physicians India 41: 94–96PubMedGoogle Scholar
  32. Devey M, Bleasdale K, Lee S, Rath S (1988) Determination of the functional affinity of IgG1 and IgG4 antibodies to tetanus toxoid by isotype-specific solid-phase assays. J Immunol Methods 106: 119–125PubMedGoogle Scholar
  33. Dezfulian M, Bitar RA, Bartlett JG (1987) Kinetics study of immunological response to Clostridium botulinum toxin. J Clin Microbiol 25: 1336–1337PubMedGoogle Scholar
  34. Dimache G, Stoean C, Durbaca S, Croitoru M, Ionescu M, Nedelcu IN, Corbu I (1990) Study of specific immune response to unadsorbed concentrated tetanus vaccine administered by intradermal route to non-immunized persons in the last ten years. Arch Roum Pathol Exp Microbiol 49: 51–62PubMedGoogle Scholar
  35. Doellgast GJ, Triscott MX, Beard GA, Bottoms JD, Cheng T, Roh BH, Roman MG, Hall PA, Brown JE (1993) Sensitive enzyme-linked immunosorbent assay for detection of Clostridium botulinum neurotoxins A,B, and E using signal amplification via enzyme-linked coagulation assay. J Clin Microbiol 31: 2402–2409PubMedGoogle Scholar
  36. Doellgast GJ, Beard GA, Bottoms JD, Cheng T, Roh BH, Roman, MG, Hall PA, Triscott MX (1994a) Enzyme-linked immunosorbent assay and enzyme-linked coagulation assay for detection of Clostridium botulinum neurotoxins A,B, and E and solution-phase complexes with dual-label antibodies. J Clin. Microbiol 32: 105–111PubMedGoogle Scholar
  37. Doellgast GJ, Triscott MX, Beard GA, Bottoms JD (1994b) Enzyme-linked immunosorbent assay-enzyme-linked coagulation assay for detection of antibodies to Clostridium botulinum neurotoxins A,B, and E and solution phase complexes. J Clin Microbiol 32: 851–853PubMedGoogle Scholar
  38. Duff JT, Wright GG, Klerer J, Moore DE, Bibler RH (1957a) Studies on immunity to toxins of Clostridium botulinum. I. A simplified procedure for isolation of type A toxin. J Bacteriol 73: 42–47PubMedGoogle Scholar
  39. Duff JT, Klerer J, Bibler RH, Moore DE, Gottfried C, Wright GG (1957b) Studies on immunity to toxins of Clostridium botulinum. II. Production and purification of type B toxin for toxoid. J Bacteriol 73: 597–601PubMedGoogle Scholar
  40. Eisel U, Jarausch W, Goretzki K, Henschen A, Engels J, Weiler U, Hudel M, Habermann Nieman H (1986) Tetanus toxin: primary structure, expression in E. coli, and homology with botulinum toxins. EMBO J 10: 2495–2502Google Scholar
  41. Englund JA, Mbawuike IN, Hammill H, Holleman MC, Baxter BD, Glezen WP (1993) Maternal immunization with influenza or tetanus toxoid vaccine for passive antibody protection in young infants. J Infect Dis 168: 647–656PubMedGoogle Scholar
  42. Fairweather NF, Lyness VA (1986) The complete nucleotide sequence of tetanus toxin. Nucleic Acid Res 14: 7809–7812PubMedGoogle Scholar
  43. Fairweather NF, Lyness VA, Pickard DJ, Allen G, Thomson RO (1986) Cloning, nucleotide sequencing, and expression of tetanus toxin fragment C in Escherichia coli. J Bacteriol 165: 21–27PubMedGoogle Scholar
  44. Fairweather NF, Lyness VA, Maskell DJ (1987) Immunization of mice against tetanus with fragments of tetanus toxin synthesized in Escherichia coli. Infect Immun 55: 2541–2545PubMedGoogle Scholar
  45. Fairweather NF, Chatfield SN, Makoff AJ, Strugnell RA, Bester J, Maskell DJ, Dougan G (1990) Oral vaccination of mice against tetanus by use of a live attenuated Salmonella carrier. Infect Immun 58: 1323–1326PubMedGoogle Scholar
  46. Finney DJ (1971) Probit analysis, 3rd edn. University Press Cambridge, CambridgeGoogle Scholar
  47. Fiock MA, Devine LF, Gearinger NF, Duff JT, Wright GG, Kadull PJ (1962) Studies on immunity to toxins of Clostridium VIII. immunological response of man to purified bivalent AB botulinum toxoid. J Immunol 88: 277–283PubMedGoogle Scholar
  48. Fiock MA, Cardella MA, Gearinger NF (1963) Studies on immunity of toxins of Clostridium botulinum. IX. Immunological response of man to purified pentavalent ABCDE botulinum toxoid. J Immunol 90: 697–702PubMedGoogle Scholar
  49. Frankovich TL, Arnon SS (1991) Clinical trial of botulism immune globulin for infant botulism. West J Med 154: 103PubMedGoogle Scholar
  50. Franz DR, Pitt LM, Clayton MA, Hanes MA, Rose KJ (1993) Efficacy of prophylactic and therapeutic administration of antotoxin for inhalation botulism. In: DasGupta BR (ed) Botulinum and tetanus neurotoxins, neurotransmission and biomedical aspects. Plenum, New York, pp 473–476Google Scholar
  51. French M, Harrison G (1985) The IgG subclasses of tetanus toxoid antibody following immunization: studies in healthy adults and patients with chronic or recurrent respiratory tract infections. J Clin Lab Immunol 18: 21–25Google Scholar
  52. Fulthorpe AJ (1955) Clostridium botulinum type A and type B toxin-antitoxin reactions. J Hyg 53: 180–195Google Scholar
  53. Gentili G, Pini C, Collotti C (1984) The determination of the potency of human tetanus immunoglobulins by an enzyme-linked immunosorbent assay. J Biol Stand 12: 167–173PubMedGoogle Scholar
  54. Gentili G, Pini C, Collotti C (1985) The use of immunoenzymatic assay for the estimation of tenanus antitoxin in human sera: a comparison with seroneutralization and indirect haemagglutination. J Biol Stand 13: 53–59PubMedGoogle Scholar
  55. Gentili G, Wirz M, Collotti C (1986) Clostridium tetani toxin antibodies. In: Masseyeff R (ed) Antigens and antibodies in chlamydial and bacterial diseases, XI. 3rd edn VCH Weinheim pp 44–52Google Scholar
  56. Gentili G, D’Amelio R, Wirz M, Matricardi PM, Nisini R, Collotti C, Pasquini P, Stroffolini T (1993) Prevalence of hyperimmunization against tetanus in Italians born after the introduction of mandatory vaccination of children with tetanus toxoid in 1968. Infection 21: 80–82PubMedGoogle Scholar
  57. German FM, Bizzini B, German A (1987a) Immunity to tetanus: tetanus antitoxin and anti-Bllb in human sera. J Biol Stand 15: 223–230PubMedGoogle Scholar
  58. German FM, German A, Bizzini B (1987b) Relation between protective potency and specificity of antibodies in sera of tetanus immunized individuals. Eur J Epidemiol 3: 119–122PubMedGoogle Scholar
  59. Gigliotti F, Insel RA (1982) Protective human hybridoma antibody to tetanus toxin. J Clin Invest 70: 1306–1309PubMedGoogle Scholar
  60. Glenny AT, Barr M (1932) The “dilution ratio” of diphtheria antitoxin as a measure of avidity. J Pathol Bacteriol 35: 91–96Google Scholar
  61. Gonnering RS (1988) Negative antibody response to long-term treatment of facial spasm with botulinum toxin. Am J Ophthalmol 105: 313–315PubMedGoogle Scholar
  62. Greene PE, Fahn S (1993) Use of botulinum toxin type F injections to treat torticollis in patients with immunity to botulinum toxin type A. Mov Disord 8: 479–483PubMedGoogle Scholar
  63. Gupta RJ, Higham S, Gupta CK, Rost BR, Siber GR (1994) Suitability of the vero cell method for titration of diphtheria antitoxin in the United States potency test for diphtheria toxoid. Biologicals 22: 65–72PubMedGoogle Scholar
  64. Habig W, Tankersley D (1991) Tetanus. In: Cryz S (ed) Vaccines and immunotherapy. Pergamon, New York, pp 13–19Google Scholar
  65. Halpern JL, Habig WH, Neale EA, Stibitz S (1990) Cloning and expression of functional fragment C of tetanus toxin. Infect Immun 58: 1004–1009PubMedGoogle Scholar
  66. Hambleton P, Cohen HE, Palmer BJ, Melling J (1992) Antitoxins and botulinum toxin treatment. BMJ 304: 959–960PubMedGoogle Scholar
  67. Harrell WK, Green JH, Winn JF (1964) Preparation, evaluation, and use of C. botulinum antitoxins. In: Lewis KH, Cassel KJ (eds) Botulism. US Department of Health, Education, and Welfare, Cincinnati, pp 165–170Google Scholar
  68. Hatheway CL, Dang C (1994) Immunogenicity of the neurotoxins of Clostridium botulinum. In: Jankovoc J, Hallett M (ed) Therapy with botulinum toxins. Dekker, New York, pp 93–107Google Scholar
  69. Hatheway CH, Snyder JD, Seals JE, Edell TA, Lewis GJ (1984) Antitoxin levels in botulism patients treated with trivalent equine botulism antitoxin to toxin types A,B, and E. J Infect Dis. 150: 407–412PubMedGoogle Scholar
  70. Hazlewood M, Nusrat R, Kumararatne DS, Goodall M, Raykundalia C, Wang DG, Joyce HJ, Milford WA, Forte M, Pahor A (1993) The acquisition of anti-pneumococcal capsular polysaccharide Haemophilus influenzae type b and tetanus toxoid antibodies, with age, in the UK. Clin Exp Immunol 93: 157–164PubMedGoogle Scholar
  71. Helting TB, Zwisler O (1977) Structure of tetanus toxin I. Breakdown of the toxin molecule and discrimination between polypeptide fragments. J Biol Chem 252: 187–193PubMedGoogle Scholar
  72. Hendriksen CFM, vander Gun JW, Nagel J, Kreeftenberg JG (1988) The toxin binding inhibition test as a reliable in vitro alternative to the toxin neutralization test in mice for the estimation of tetanus antitoxin in human sera. J Biol Stand 16: 287–297PubMedGoogle Scholar
  73. Hendriksen CFM, vander Gun JW, Kreeftenberg JG (1989) Combined estimation of tetanus and diphtheria antitoxin in human sera by the in vitro toxin-binding inhibition (ToBI) test. J Biol Stand 17: 191–200PubMedGoogle Scholar
  74. Hendriksen CFM, vander Gun JW, Marsman F, Kreeftenberg JG (1991) The use of the in vitro toxin binding (ToBI) test for the estimation of the potency of tetanus toxoid. Biologicals 19: 23–29PubMedGoogle Scholar
  75. Hodgson J (1991) Making monoclonals in microbes. Bio technol 9: 421–425Google Scholar
  76. Jackson RJ, Fujihashi K, Xu-Amano J, Kiyono H, Elson CO, McGhee JR (1993) Optimizing oral vaccines: induction of systemic and mucosal B-cell and antibody responses to tetanus toxoid by use of cholera toxin as an adjuvant. Infect Immun 61: 4272–4279PubMedGoogle Scholar
  77. Jankovic J, Schwartz KS (1991) Clinical correlates of response to botulinum toxin injections. Arch Neurol 48: 1253–1256PubMedGoogle Scholar
  78. Jerne NK (1951) Avidity. Acta Pathol. Microbiol Scand. [Suppl] 87.Google Scholar
  79. Jubb TF, Ellis TM, Gregory AR (1993) Diagnosis of botulism in cattle using ELISA to detect antibody to botulinum toxins. Aust Vet J 70: 226–227PubMedGoogle Scholar
  80. Kaiser II, Middlebrook JL (1988) Preparation of a crotoxin neutralizing monoclonal antibody. Toxicon 26: 855–865PubMedGoogle Scholar
  81. Kamei M, Hashizume S, Sugimoto N, Ozutsumi K, Matsuda M (1990) Establishment of stable human/mouse hybrid cell lines producing large amounts of anti-tetanus human monoclonal antibodies with high neutralizing activity. Eur J Epidemiol 6: 386–397PubMedGoogle Scholar
  82. Kempner W (1897) Weitere Beiträge zur Lehre der Fleischvergiftung. Das Antitoxin des Botulismus. Z. Hyg. Infektkr 26: 418–500Google Scholar
  83. Kenimer JG, Habig WH, Hardegree MC (1983) Monoclonal antibodies as probes of tetanus toxin structure and function. Infect Immun 42: 942–948PubMedGoogle Scholar
  84. Kiessig ST, Hentschel C, Jahn S, Mehl M. Starke R, Porstmann T (1991) A solid-phase enzyme immunoassay for the detection of tetanus toxin using human and murine monoclonal antibodies. J Basic Microbiol 31: 135–140PubMedGoogle Scholar
  85. Krag P, Bentzon MW (1965) Report on international collaborative assay of antibotulinus serum type F. WHO/BS 750: 65 (unpublished)Google Scholar
  86. Lang AB, Cryz SJ Jr, Schurch U, Ganss MT, Bruderer U (1993) Immunotherapy with human monoclonal antibodies. Fragment A specificity of polyclonal and monoclonal antibodies is crucial for full protection against tetanus toxin. J Immunol 151: 466–472PubMedGoogle Scholar
  87. LaPenotiere HF, Clayton MA, Brown JE, Middlebrook JL (1993) Development of a molecularly engineered vaccine for C. botulinum neurotoxins. In: DasGupta BR (ed) Botulinum and tetanus neurotoxins, neurotransmission and biomedical aspects. Plenum, New York, pp 463–466Google Scholar
  88. Larrick JW, Truitt KE, Raubitschek AA, Senyk G, Wang JCN (1983) Characterization of human hybridomas secreting antibody to tetanus toxoid. Proc Natl Acad Sci USA 80: 6376–6380PubMedGoogle Scholar
  89. Layton G (1980) A microenzyme-linked immunosorbent assay (ELISA) and radioimmunosorbent technique (RIST) for the detection of immunity to clinical tetanus. Med Lab Sci 37: 323–329PubMedGoogle Scholar
  90. Layton LL, Arimoto L, Lamanna C, Olson R, Sharp D, Kondo H, Sakaguchi G (1972) Immunochemical and physiological comparison of horse botulinal antitoxins. Jpn J Med Sci Biol 25: 309–320PubMedGoogle Scholar
  91. Lee HF, Tseng LR (1991) Improvement of the E1A kit for the detection of tetanus immunoglobulin in human sera. Chung Hua Min Kuo Wei Sheng Wu Chi Mien I Hsueh Tsa Chih 24: 221–228PubMedGoogle Scholar
  92. Lee HF, Chen HM, Hsu ST (1987) Serological survey of immune response to tetanus toxoid using antitoxin enzyme immunoassay. Chung Hua Min Kuo Wei Sheng Wu Chi Mien I Hsueh Tsa Chih 20: 269–278PubMedGoogle Scholar
  93. Leuchs J (1910) Beiträge zur Kenntnis des Toxins and Antitoxins des Bacillus botulinus. Z Hyg Infektkr 65: 55–84Google Scholar
  94. Lewis GE (1981) Approaches to the prophylaxis, immunotherapy, and chemotherapy of botulism. In: Lewis G E Jr (ed) Biomedical aspects of botulism. Academic, New York, pp 261–270Google Scholar
  95. Lewis GE Jr, Metzger JF (1978) Botulism immune plasma. Lancet 2: 634–635PubMedGoogle Scholar
  96. Lewis GE Jr, Metzger JF (1980) Studies on the Prophylaxis and treatment of botulism. In: Eaker D, Wadstrom T (eds) Natural toxins, Pergamon, Oxford, pp 601–606Google Scholar
  97. Luisto M (1990) Tetanus in Finland: diagnostic problems and complications. Ann Med 22: 15–19PubMedGoogle Scholar
  98. Lyng J, Heron I (1992) Quantitative estimation of diphtheria and tetanus toxoids. VI. Use of different antibody titration methods for evaluation of immunogenicity in animals during potency assay of diphtheria toxoid. Biologicals. 20: 101–108PubMedGoogle Scholar
  99. Makoff AJ, Ballantine SP, Smallwood AE, Fairweather NF (1989a) Expression of tetanus toxin fragment C in E. coli: its purification and potential use as a vaccine. Biotechnol 7: 1043–1046Google Scholar
  100. V. Chandrika CTMI Volume 195 Chapter 5 Mss 213 toGoogle Scholar
  101. Makoff AJ, Oxer MD, Romanos MA, Fairweather NF, Ballantine S (1989b) Expression of tetanus toxin fragment C in E. coli: high level expression by removing rare codons. Nucleic Acids Res 17: 10191–10202PubMedGoogle Scholar
  102. Markela E, Stalhberg TH, Hemmilä I (1993) Europium-labeled recombinant protein G. A fast and sensitive universal immunoreagent for time-resolved immunofluorometry. J Immunol Methods 161: 1–6PubMedGoogle Scholar
  103. Maskell DJ, Sweeney KJ, O’Callaghan D, Hormaeche CE, Liew FY, Dougan G (1987) Salmonella typhimuriam aroA mutants as carriers of the Escherichia coli heat-labile enterotoxin to the murine secretory and systemic immune systems. Microb Pathog 2: 211–221PubMedGoogle Scholar
  104. Mastroeni P, Leonardi MS, Gazzara D, Bizzini B (1989) Rapid assessment of the antitetanus immune status of a subject using dot-ELISA. Eur J Epidemiol 5: 97–100PubMedGoogle Scholar
  105. Matsuda M, Yoneda M (1975) Isolation and purification of two antigenically active, “complementary” polypeptide fragments of tetanus toxin. Infect Immun 12: 1147–1153PubMedGoogle Scholar
  106. McComb J (1964) The prophylactic dose of homologous tetanus antitoxin. N Engl J Med 270: 175–178PubMedGoogle Scholar
  107. McCracken GH Jr, Dowell DL, Marshall FN (1971) Double-blind trial of equine antitoxin and human immune globulin in tetanus neonatorum. Lancet 1: 1146–1149PubMedGoogle Scholar
  108. Melville-Smith M (1990) Diphtheria and tetanus antitoxins. In: Wreghitt TG, Morgan-Capner P (eds) ELISA in the clinical microbiology laboratory. Public Health Laboratory Service, London, pp 136–147Google Scholar
  109. Metzger JF, Lewis GE (1979) Human-derived immune globulins for the treatment of botulism. Rev Infect Dis 1: 689–692PubMedGoogle Scholar
  110. Middlebrook JL (1991) Preparation and characterization of monoclonal antibodies against pseudexin. Toxicon 29: 359–370PubMedGoogle Scholar
  111. Moen RC, Oemichen SL, Kiggnes AJ, Hong R (1986) ELISA detection of specific functional antibodies in human sera to Escherichia coli, tetanus toxoid, and diphtheria-tetanus toxoids: normal values for IgG, IgA, and IgM. Diagn Immunol 4: 17–23PubMedGoogle Scholar
  112. Mouillet L, Chaniot S, Crajer M, Netter R (1978) Intérêt de l’immunodiffusion radiale dans le titrage des immunoglobulines tétaniques. J Biol Stand 6: 13–20Google Scholar
  113. Nation NS, Pierce NF, Adler SJ, Chinnock RF, Wehrle PF (1963) Tetanus: use of human hyperimmune globulin in treatment. Calif Med 98: 305–307PubMedGoogle Scholar
  114. Niemann H (1992) Clostridial neurotoxins-proposal of a common nomenclature. Toxicon 30: 223–225PubMedGoogle Scholar
  115. Nyerges G, Lutter J (1980) The influence of the method of preservation of erythrocytes on the correlation between tetanus antitoxin values as measured by passive haemagglutination and by sero-neutralization tests in mice. J Biol Stand 8: 311–316PubMedGoogle Scholar
  116. Oberst FW, Crook JW, Cresthull P, House MJ (1967) Evaluation of botulinum antitoxin, supportive therapy, and artificial respiration in monkeys with experimental botulism. Clin Pharmacol Ther 9: 209–214Google Scholar
  117. Ohishi I, Sakaguchi G, Riemann H, Behymer D, Hurvell B (1979) Antibodies to Clostridium botulinum toxins in free-living birds and mammals. J Wild Dis 15: 3–9Google Scholar
  118. Pal A, Kumar R, Jailkhani BL (1990) An ELISA for quantitation of tetanus toxin. Indian J Med Res 91: 124–125PubMedGoogle Scholar
  119. Paul-Murphy J, Gershwin LJ, Thatcher EF, Fowler ME, Habig WH (1989) Immune response of the llama (Lama glama) to tetanus toxoid vaccination. Am J Vet Res 50: 1279–1281PubMedGoogle Scholar
  120. Pearce LB, Borodic GE, Johnson EA, First ER, MacCallum R (1994a) The median paralysis unit: a more pharmacologically relevant unit of biologic activity for botulinum toxin. Toxicon (submitted)Google Scholar
  121. Pearce LB, Borodic GE, First ER, MacCallum R (1994b) Measurement of botulinum toxin activity: evaluation of the lethality assay. Ther Appl Pharmacol (in press)Google Scholar
  122. Peel M (1980a) Measurement of tetanus antitoxin. I. Indirect haemagglutination. J Biol Stand 8: 177–189PubMedGoogle Scholar
  123. Peel M (1980b) Measurement of tetanus antitoxin. II. Toxin neutralization. J Biol Stand 8: 191–207PubMedGoogle Scholar
  124. Porter JD, Parkin MA, Corbel MJ, Farrington CP, Watkins JT, Begg NT (1992) Lack of early antitoxin response to tetanus booster. Vaccine 10: 334–336PubMedGoogle Scholar
  125. Prevots R, Sutter RW, Strebel PM, Cochi SL, Hadler S (1992) Tetanus surveillance—United States, 1989–1990. MMRW CDC Surveill Summ 41: 1–9Google Scholar
  126. Ray P, Berman JD, Middleton W, Brendle J (1993) Botulinum toxin inhibits arachidonic acid release associated with acetylcholine release from PC12 cells. J Biol Chem 268: 11057–11064PubMedGoogle Scholar
  127. Reames HR, Kadull PJ, Housewright RD, Wilson JB (1947) Studies on botulinum toxoids, types A and B. III. Immunization in man. J Bacteriol 55: 309–324Google Scholar
  128. Reed LJ, Muench HM (1938) A simple method for estimating fifty percent endpoints. Am J Hyg 27: 493–497Google Scholar
  129. Reiss J, Zawistowska MI, Grzybowski J (1991) A simplified method of rapid hemagglutination test for determining the levels of tetanus antibodies. Med Dosw Mikrobiol 43: 51–54PubMedGoogle Scholar
  130. Rey M, Fontanges R, Robert D, Taylor J, Relyveld E, Vincent-Falquet J, Durand B (1978) Le test d’hémagglutination passive dans l’évaluation de l’immunité antitétanique. Dev Biol Stand 41: 55–63PubMedGoogle Scholar
  131. Romanos MA, Makoff AJ, Fairweather NF, Beesley KM, Slater DE, Rayment FB, Payne MM, Clare JJ (1991) Expression of tetanus toxin fragment C in yeast: gene synthesis is required to eliminate fortuitous polyadenylation sites in AT-rich DNA. Nucleic Acids Res 19: 1461–1467PubMedGoogle Scholar
  132. Rubin LG, Dezfulian M, Yolken RH (1982) Serum antibody response to Clostridium botulinum toxin in infant botulism. J Clin Microbiol 16: 770–771PubMedGoogle Scholar
  133. Rubin M, Wong E, Bell H, Bowman J, Albritton W (1981) A reversed rocket electrophoresis method for the determination of high tetanus antitoxin levels. J Biol Stand 9: 367–371PubMedGoogle Scholar
  134. Rumke HC, Schlumberger M, Floury B, Nagel J, van Steenis B (1993) Serological evaluation of a simplified immunization schedule using quadruple DPT-polio vaccine in Burkina Faso. Vaccine 11: 1113–1118PubMedGoogle Scholar
  135. Schroder JP, Kuhlmann WD (1991) Detection of tetanus antitoxin using Eu(3+)- labeled anti-human immunoglobulin G monoclonal antibodies in a time-resolved fluorescence immunoassay. J Clin Microbiol 29: 1504–1507PubMedGoogle Scholar
  136. Schwarz PJ, Arnon SS (1992) Botulism immune globulin for infant botulism arrives—one year and a Gulf War later. West J Med 156: 197–198PubMedGoogle Scholar
  137. Sesardic D, Corbel MJ (1992) Testing for neutralising potential of serum antibodies to tetanus and diphtheria toxin (letter). Lancet 340: 737–738PubMedGoogle Scholar
  138. Sesardic D, Wong MY, Gaines Das RE, Corbell MJ (1994) The 1st international standard for antitetanus immunoglobulin, human; pharmaceutical, evaluation and international collaborative study. Biologicals 21: 67–75Google Scholar
  139. Sheppard AJ, Hughes M, Stephen J (1987) Affinity purification of tetanus toxin using polyclonal and monoclonal antibody immunoadsorbents. J Appl Bacteriol 62: 335–348PubMedGoogle Scholar
  140. Shone CC, Melling J (1992) Inhibition of calcium-dependent release of noradrenaline from PC12 cells by botulinum type-A neurotoxin. Long-term effects of the neurotoxin on intact cells. Eur J Biochem 207: 1009–1016PubMedGoogle Scholar
  141. Shone C, Appleton N, Wilton SP, Hambleton P, Modi N, Gatley S, Melling J (1986) In vitro assays for botulinum toxin and antitoxins. Dev Biol Stand 64: 141–145PubMedGoogle Scholar
  142. Siatkowski RM, Tyutyunikov A, Biglan AW, Scalise D, Genovese C, Raikow RB, Kennerdell JS, Feuer WJ (1993) Serum antibody production to botulinum A toxin. Ophthalmology 100: 1861–1866PubMedGoogle Scholar
  143. Siegel LS (1988) Human immune response to botulinum pentavalent (ABCDE) toxoid determined by a neutralization test and by an enzyme-linked immunosorbent assay. J Clin Microbiol 26: 2351–2356PubMedGoogle Scholar
  144. Siegel LS (1989) Evaluation of neutralizing antibodies to type A, B, E, and F botulinum toxins in sera from human recipients of botulinum pentavalent (ABCDE) toxoid. J Clin Microbiol 27: 1906–1908PubMedGoogle Scholar
  145. Simonsen O (1989) Vaccination against tetanus and diphtheria. Evaluation of immunity in the Danish population, guidelines for revaccination, and methods for control of vaccination programs. Dan Med Bull 36: 24–47PubMedGoogle Scholar
  146. Simonsen O, Kjeldsen K, Heron I (1984) Immunity against tetanus and effect of revaccination 25–30 years after primary vaccination. Lancet 2: 1240–1242PubMedGoogle Scholar
  147. Simonsen O, Bentzon MW, Heron I (1986) ELISA for the routine determination of antitoxic immunity to tetanus. J Biol Stand 14: 231–239PubMedGoogle Scholar
  148. Simonsen O, Schou C, Heron I (1987) Modification of the ELISA for the estimation of tetanus antitoxin in human sera. J Biol Stand 15: 143–157PubMedGoogle Scholar
  149. Simpson LL, Lake P, Kozaki S (1990) Isolation and characterization of a novel human monoclonal antibody that neutralizes tetanus toxin. J Pharmacol Exp Ther 254: 98–103PubMedGoogle Scholar
  150. Sjogren MH, McKee KT, Clayton MA, Franz DR, Condie RM, Peters CJ (1993) Safety and kinetics of an equine F(ab)2 heptavalent botulinum antitoxin in human volunteers. In: Kamely D, Bannister KA, Sasmor RM (eds) Army Science: the new frontiers: military and civilian applications. Borg Biomedical, Saratoga, WY, pp 157–158Google Scholar
  151. Stevens RH, Saxon A (1978) Immunoregulation in humans: control of antitetanus toxoid antibody production after booster immunization. J Clin Invest 62: 1154–1160PubMedGoogle Scholar
  152. Stiffler-Rosenberg G, Fey H (1977) Measurement of human tetanus antitoxinusing an enzyme-linked immunosorbent assay. Schweiz Med Wochenschr. 107:Google Scholar
  153. Stover CK, de la Cruz VF, Fuerst TR, Burlein JE, Benson LA, Bennett LT, Bansal GP, Young JF, Lee MH, Hatfull GF, Snapper SB, Barletta RG, Jacobs WR Jr, Bloom BR (1991) New use of BCG for recombinant vaccines. Nature 351: 456–460PubMedGoogle Scholar
  154. Sugiyama H, Brenner SL, DasGupta BR (1975) Detection of Clostridium botulinum toxin by local paralysis elicited with intramuscular challenge. Appl Microbiol 30: 420–423PubMedGoogle Scholar
  155. Tacket CO, Shandera WX, Mann JM, Hargrett NT, Blake PA (1984) Equine antitoxin use and other factors that predict outcome in type A foodborne botulism. Am J Med 76: 794–798PubMedGoogle Scholar
  156. Takahashi M, Komiya T, Kameyama S, Sakaguchi G (1990a) Titration of botulinum antitoxin of low levels by the score method. Jpn J Med Sci Biol 43: 163–170PubMedGoogle Scholar
  157. Takahashi M, Noda H, Takeshita S, Fujiwara T, Nakanoin H, Mizunoya T, Sakaguchi G (1990b) Attempts to quantity Clostridium botulinum type A toxin and antitoxin in serum of two cases of infant botulism in Japan. Jpn J Med Sci Biol 43: 233–237PubMedGoogle Scholar
  158. Thiele GM, Rogers J, Collins M, Yasuda N, Smith D, McDonald TL (1990) An enzyme-linked immunosorbent assay for the detection of antitetanus toxoid antibody using aluminium-absorbed coating antigen. J Clin Lab Anal 4: 126–129PubMedGoogle Scholar
  159. Thompson DE, Brehm JK, Oultram JD, Swinfield T-J, Shone CC, Atkinson T, Melling J, Minton NP (1990) The complete amino acid sequence of the Clostridium botulinum type A neurotoxin, deduced by nucleotide sequence analysis of the encoding gene. Eur J Biochem 189: 73–81PubMedGoogle Scholar
  160. Trabaud M-A, Lery L, Desgranges C (1989) Human monoclonal antibodies with a protective activity against tetanus toxin. Acta Pathol Microbiol Immunol Scand 97: 671–676Google Scholar
  161. Velikanov IM (1934) Experimental immunization of man against botulism. Klin Med 12: 1802–1806Google Scholar
  162. Vernacchio L, Madico G, Verastegui M, Diaz F, Collins TS, Gilman RH (1993) Neonatal tetanus in Peru: risk assessment with modified enzyme-linked immunosorbent assay and toxoid skin test. Am J Public Health 83: 1754–1756PubMedGoogle Scholar
  163. Virella G, Fudenberg H, Kyong K, Pandey J, Galbriath R (1978) A practical method for quantitation of antibody responses to tetanus and diphtheria toxoids in normal young children. Z Immunitätsforsch Immunobiol 155: 80–86PubMedGoogle Scholar
  164. Virella G, Hyman B, Kilpatrick JM (1985) Solid-phase enzymoimmunoassay of antitetanus toxoid antibodies. Diagn Immunol 3: 215–219PubMedGoogle Scholar
  165. Volk WA, Bizzini B, Snyder RM, Bernhard E, Wagner RR (1984) Neutralization of tetanus toxin by distinct monoclonal antibodies binding to multiple epitopes on the toxin molecule. Infect Immun 45: 604–609PubMedGoogle Scholar
  166. Weber JT, Hibbs RJ, Darwish A, Mishu B, Corwin AL, Rakha M, Hatheway CL, El-Sharkawy SS, El- Rahim SA, Al-Hamd FS, Sam JE, Blake PA, Tauxe RV (1993) A massive outbreak of type E botulism associated with traditional salted fish in Cairo. J Infect Dis 167: 451–454PubMedGoogle Scholar
  167. Wells JM, Wilson PW, Norton PM, Gasson MJ, LePage RWF (1993) Lactococcus lactis: high-level expression of tetanus toxin fragment C and protection against lethal challenge. Mol Microbiol 8: 1155–1162PubMedGoogle Scholar
  168. WHO (1993) Expanded program on immunization. The global elimination of neonatal tetanus: progress to date. Wkly Epidemiol Rec 68: 277–282Google Scholar
  169. WHO (1977) Manual for the production and control of vaccines: tetanus toxoid. BLG/UNDP/77.2 Rev 1. WHO, GenevaGoogle Scholar
  170. WHO Expert Committee on Biological Standardization (1982) Clostridium botulinum Type B Antitoxin: replacement of the international standard progress report. WHO/BS 82. 1352 (unpublished)Google Scholar
  171. Wilde H, Chutivongse S (1990) Equine rabies immune globulin: a product with an undeserved poor reputation. Am J Trop Med Hyg 42: 175–178PubMedGoogle Scholar
  172. Wilde H, Chomchey P, Prakongsri S, Punyaratabandhu P (1987) Safety of equine rabies immune globulin (letter) Lancet 2: 1275PubMedGoogle Scholar
  173. Wilde H, Chomchey P, Prakongsri S, Punyaratabandhu P, Chutivongse S (1989a) Adverse effects of equine rabies immune globulin. Vaccine 7: 10–11PubMedGoogle Scholar
  174. Wilde H, Chomchey P, Punyaratabandhu P, Phanupak P, Chutivongse S (1989b) Purified equine rabies immune globulin: a safe and affordable alternative to human rabies immune globulin. Bull World Health Organ 67: 731–736PubMedGoogle Scholar
  175. Williamson LC, Fitzgerald SC, Neale EA (1992) Differential effects of tetanus toxin on inhibitory and excitatory neurotransmitter release from mammalian spinal cord cultures in culture. J Neurochem 59: 2148–2157PubMedGoogle Scholar
  176. Wirz M, Gentili G, Collotti C (1990) Tetanus vaccine: present status. Adv Biotechnol Processes 13: 35–55PubMedGoogle Scholar
  177. Wright GG, Duff JT, Fiock MA, Devlin HB, Söderström RL (1960) Studies on immunity to toxins of Clostridium botulinum. V.Detoxification of purified type A and B toxins, and the antigenicity of univalent and bivalent aluminum phosphate adsorbed toxoids. J Immunol 84: 384–389PubMedGoogle Scholar
  178. Ziegler-Heitbrock HWL, Reiter C, Trenkmann J, Futterer A, Reithmuller G (1986) Protection of mice against tetanus toxin by combination of two human monoclonal antibodies recognizing distinct epitopes on the toxin molecule. Hybridoma 5: 21–31PubMedGoogle Scholar
  179. Zuber M, Sebald M, Bathien N, de Recondo RJ, Rondot P (1993) Botulinum antibodies in dystonic patients treated with type A botulinum toxin: frequency and significance Neurology 43: 1715–1718PubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1995

Authors and Affiliations

  • J. L. Middlebrook
    • 1
  • J. E. Brown
    • 1
  1. 1.Toxinology DivisionU.S. Army Medical Research Institute of Infectious DiseasesFrederickUSA

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